Regeneration in the adult central nervous system (CNS) is thought to be hampered by the lesion-induced activation of astrocytes and meningeal cells and the consecutive formation of a glial scar. The substrate properties of reactive astrocytes differ significantly from their neonatal counterparts, which promote axon growth, but in spite of intensive studies the underlying molecular changes are still not fully understood. We have used two cell culture systems to compare the expression of certain surface molecules on neonatal astrocytes, reactive astrocytes and meningeal cells in vitro. Both, neonatal and reactive adult astrocytes exhibited a very similar expression of growth promoting molecules (NCAM, L1, laminin, fibronectin, DSD-1 proteoglycan) and potential inhibitors (tenascinC, chondroitin sulfate, and NG2-proteoglycan), whereas we could not detect the inhibitory keratan sulfate on either astrocyte population. In contrast, meningeal cells expressed considerable levels of keratan sulfate, but only minimal amounts of NCAM. In addition, the much higher expression of extracellular fibronectin around meningeal cells implies an excess formation of extracellular matrix (ECM). In coculture experiments, embryonic retinal ganglion cell (RGC) axons clearly avoided meningeal cells and instead preferred even reactive adult astrocytes. Our results suggest that the expression of inhibitory keratan sulfate proteoglycans together with a lack of NCAM and an excess production of ECM may be responsible for the non-permissiveness of meningeal cells. Compared to reactive astrocytes, meningeal cells are even worse a substrate for growing axons. None of the molecules investigated, however, seems to account for the different substrate properties of neonatal and reactive adult astrocytes.